Pumpkin seed oil is well known for its health benefits due to its high content of fatty acid constituents and tocopherols. The removal efficacy of pumpkin seed oil was assessed using UV–Vis spectroscopy. The oil was able to remove 79.92 ± 0.07%, 41.02 ± 0.25%, and 23.54 ± 0.19% of foundation and liquid and pen eyeliners. A stable makeup remover was formulated using 5–15% pumpkin seed oil. Addition of pumpkin seed oil significantly (P < 0.001) enhanced removal ability of the base remover. The remover containing 5% pumpkin seed oil was able to remove 89.27 ± 0.02%, 67.72 ± 0.08%, and 41.25 ± 0.07% of foundation, liquid, and pen eyeliners, respectively, while those of the remover containing 10% pumpkin seed oil were 78.24 ± 0.02%, 66.88 ± 0.05%, and 38.43 ± 0.05%, and those of the remover containing 15% pumpkin seed oil were 84.41 ± 0.01%, 69.79 ± 0.12%, and 41.88 ± 0.04%, respectively. On the other hand, removal efficiencies of the benchmark were 91.20 ± 0.03%, 73.46 ± 0.10%, and 54.00 ± 0.07%, respectively. The removers containing pumpkin seed oil did not cause skin irritation as monitored by a single closed‐patch test in 10 female volunteers. The remover containing 5% pumpkin seed oil was further preference studied in 25 female volunteers in a comparison with the benchmark. The pumpkin seed oil remover gained a better overall preference over the benchmark (82.29 ± 4.17% and 80.20 ± 8.64%; P = 0.287). Of which, skin hydration of the developed bio‐oil remover was significantly (P < 0.001) satisfied.